Herbimycin A blocks IL‐1‐induced NF‐κB DNA‐binding activity in lymphoid cell lines

Iwasaki, Takaaki; Uehara, Yoshimasa; Graves, Lee M.; Rachie, N A; Bomsztyk, Karol

doi:10.1016/0014-5793(92)80067-q

Cited by 66 publications

(22 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The ability of herbimycin A to inhibit the activation of NF-B by PMA was in direct contrast to a previous study (20), which reported that PMA activation of NF-B in 70Z/3 pre-B cells was insensitive to the drug. 70Z/3 cells differ from EL4 cells in that the activation of NF-B by PMA in the cells is more rapid.…”

Section: Herbimycin a Inhibits Nf-b Activation Induced By Il-1 And Pmcontrasting

confidence: 54%

“…The evidence, however, has been largely circumstantial, with data being presented for tyrosine kinase changes in response to IL-1 that were inhibited by herbimycin A or another tyrosine kinase inhibitor, genistein (20,21). These results were then used as evidence for tyrosine kinase involvement in the activation of NF-B, which also proved susceptible to inhibition by the compounds.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Studies into the Effect of the Tyrosine Kinase Inhibitor Herbimycin A on NF-κB Activation in T Lymphocytes EVIDENCE FOR COVALENT MODIFICATION OF THE p50 SUBUNIT

Mahon¹,

O’Neill²

1995

Journal of Biological Chemistry

111

View full text Add to dashboard Cite

The tyrosine kinase inhibitor herbimycin A was found to block NF-B stimulation in response to interleukin-1 and phorbol 12-myristate 13-acetate in EL4.NOB-1 thymoma cells and phorbol 12-myristate 13-acetate in Jurkat T lymphoma cells. The effect appeared not to involve inhibition of tyrosine kinase activation as neither interleukin-1 nor phorbol 12-myristate 13-acetate induced major changes in tyrosine phosphorylation in EL4.NOB-1 or Jurkat cells, respectively. Herbimycin A did not interfere with IB-␣ degradation, and in unstimulated cells, it modified NF-B prior to chemical dissociation with sodium deoxycholate. Because herbimycin A is thiol-reactive, we suspected that the target was the p50 subunit of NF-B, which has a key thiol at cysteine 62. Herbimycin A inhibited DNA binding when added to nuclear extracts prepared from stimulated cells, which were shown to contain high levels of p50. Incubation of herbimycin A with 2-mercaptoethanol attenuated the effect. Herbimycin A was also shown to react directly with p50, blocking its ability to bind to the NF-B consensus sequence. However, a mutant form of p50 in which cysteine 62 was mutated to serine was insensitive to herbimycin A. Finally, we demonstrated that the compound inhibited the expression of interleukin-2 (an NF-B-regulated gene) in EL4.NOB-1 cells. These data therefore suggest that herbimycin A inhibits NF-B by modifying the p50 subunit on cysteine 62 in the NF-B complex, which blocks DNA binding and NF-B-driven gene expression. The results urge caution in the use of herbimycin A as a specific tyrosine kinase inhibitor and suggest that the development of agents that selectively modify p50 may have potential as a means of inhibiting NF-B-dependent gene transcription.

show abstract

Section: Herbimycin a Inhibits Nf-b Activation Induced By Il-1 And Pmcontrasting

confidence: 54%

Section: Discussionmentioning

confidence: 99%

Studies into the Effect of the Tyrosine Kinase Inhibitor Herbimycin A on NF-κB Activation in T Lymphocytes EVIDENCE FOR COVALENT MODIFICATION OF THE p50 SUBUNIT

Mahon¹,

O’Neill²

1995

Journal of Biological Chemistry

111

View full text Add to dashboard Cite

show abstract

“…10 -11). Given the fact that IL-1 does activate protein-tyrosine kinases (33,34), recruitment of this class of kinase to the SH3-binding domains may provide a mechanism by which KPK (17), which is engaged by a contiguous domain, is activated in response to IL-1 (Fig. 11).…”

Section: Discussionmentioning

confidence: 99%

“…Bound to the K Protein-Treatment of cells with IL-1 can result in the activation of protein-tyrosine kinases (32)(33)(34). We have previously provided evidence that KPK is activated by phosphorylation (17).…”

Section: Kpk Can Be Activated In Vitro By C-src That Is Concurrentlymentioning

confidence: 99%

The K Protein Domain That Recruits the Interleukin 1-responsive K Protein Kinase Lies Adjacent to a Cluster of c-Src and Vav SH3-binding Sites

Seuningen

Ostrowski

Bustelo

et al. 1995

Journal of Biological Chemistry

Self Cite

108

128

View full text Add to dashboard Cite

The heterogeneous ribonucleoprotein particle (hnRNP) K protein interacts with multiple molecular partners including DNA, RNA, serine/threonine, and tyrosine kinases and the product of the proto-oncogene, Vav. The K protein is phosphorylated in vivo and in vitro on serine/ threonine residues by an interleukin 1 (IL-1)-responsive kinase with which it forms a complex. In this study we set out to map the K protein domains that bind kinases. We demonstrate that the K protein contains a cluster of at least three SH3-binding sites (P1, PPGRGGRPMP-PSRR, amino acids 265-278; P2, PRRGPPPPPPGRG, 285-297; and P3, RARNLPLPPPPPPRGG, 303-318) and that each one of these sites is capable of selectively engaging c-Src and Vav SH3 domains but not SH3 domains of Abl, p85 phosphatidylinositol 3-kinase, Grb-2, and Csk. We demonstrate that the K protein domain that recruits and is phosphorylated in an RNA-dependent manner by the IL-1-responsive kinase, designated KPK for K protein kinase, is contained within the 338 -425-amino acid stretch and thus is contiguous but does not include the cluster of the SH3-binding sites. K protein and KPK co-immunoprecipitate from cell extracts with either c-Src or Vav, suggesting that K protein-KPK-c-Src and K protein-KPK-Vav complexes exist in vivo. Furthermore, in the context of K protein, c-Src can reactivate KPK in vitro. The succession of kinase-binding sites contained within the K protein that allow it to form multienzyme complexes and facilitate kinase cross-talk suggest that K protein may serve as a docking platform that promotes molecular interactions occurring during signal transduction.

show abstract

“…5). NF-icB bands are readily detectable in gels of nuclear extracts prepared from PMA or TNF-a-stimulated cells; however, these bands are missing in gels of extracts from cells preincubated with herbimycin A (4 hr, 10 pg/ml) prior to stimulation (22,23).…”

mentioning

confidence: 99%

Separation of oxidant-initiated and redox-regulated steps in the NF-kappa B signal transduction pathway.

Anderson

Staal²,

Gitler³

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

361

180

View full text Add to dashboard Cite

Studies presented here show that overall NF-acB signal transduction begins with a parallel series of stimuli-specific pathways through which cytokines (tumor necrosis factor a), oxidants (hydrogen peroxide and mitomycin C), and phorbol ester (phorbol 12-myrisate 13-acetate) individually initiate ignaling. These initial pathways culminate In a common pathway through which all of the smulating agents ultimately signal NF-udl is required for NF-ucB activatio by all stimuli tested in both cell lines. Since internal sites of oxidant production have been shown to play a key role in the cytokine-stimulated activation of NF-ucB, and since t kinase and phosphatase activities are known to be altered by oxidants, these fndings suggest that intracellular redox status controls NF-dB activation by regulating tyrosine phosphorylation event(s) within the common step of the NF-ucB signal transduction pathway.The activated form of the nuclear transcription factor NF-KB increases transcription of inflammatory response genes and viral genes, notably human immunodeficiency virus (1). The unactivated form of this transcription factor is sequestered in the cytosol by member(s) of the I-4B family. T-cell stimulation by a wide variety of agents, ranging from phorbol esters (phorbol 12-myristate 13-acetate; PMA) and cytokines [tumor necrosis factor a (TNF-a) and interleukin la] to oxidants such as hydrogen peroxide (H202) and UV irradiation, cause I-KB to dissociate from the NF-KB complex. The release of NF-KB from its inhibitory protein(s) permits the translocation of NF-KB into the nucleus, where it binds to DNA enhancer motifs and regulates transcription of a diverse series ofgenes (1,2). Intracellular redox levels have been shown to regulate NF-KB signal transduction triggered by a variety of stimuli (3, 4). H202 and UV triggering of NF-#cB activation are often used as models of this common redox regulation; however, we show here that the mechanism responsible for triggering by these oxidants is genetically separable from the common, redox-sensitive mechanism that operates in response to all stimuli. We also demonstrate a common requirement for protein tyrosine phosphorylation in NF-KB activation. Since the collective activity of tyrosine kinases and phosphatases stimulated is redox regulated (5-7), we suggest that the redox regulation of these enzymes contributes to the redox sensitivity of the common step of the NF-KB signal transduction pathway.

show abstract

Herbimycin A blocks IL‐1‐induced NF‐κB DNA‐binding activity in lymphoid cell lines

Cited by 66 publications

References 31 publications

Studies into the Effect of the Tyrosine Kinase Inhibitor Herbimycin A on NF-κB Activation in T Lymphocytes EVIDENCE FOR COVALENT MODIFICATION OF THE p50 SUBUNIT

Studies into the Effect of the Tyrosine Kinase Inhibitor Herbimycin A on NF-κB Activation in T Lymphocytes EVIDENCE FOR COVALENT MODIFICATION OF THE p50 SUBUNIT

The K Protein Domain That Recruits the Interleukin 1-responsive K Protein Kinase Lies Adjacent to a Cluster of c-Src and Vav SH3-binding Sites

Separation of oxidant-initiated and redox-regulated steps in the NF-kappa B signal transduction pathway.

Contact Info

Product

Resources

About